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A gas chromatography/mass spectrometry (GC/MS) method for the quantification of nine phthalates in tattoo and permanent make-up (PMU) ink samples was developed and validated in accordance with the criteria of ISO/IEC 17025 and international guidelines. Limit of detection (LOD) and quantification (LOQ) values were 0.02 µg g−1 –0.14 µg g−1 and 0.08 µg g−1 –0.48 µg g−1, respectively. The intermediate precision, in terms of relative standard deviations (RSD), was less than 15 % over the working range selected for all the investigated phthalates. The inter-day recoveries ranged between 76 % and 116 %. The validation study demonstrated that the method is accurate and sensitive although the development of an analytical method for this matrix is quite challenging. The relative expanded uncertainty estimated with a coverage factor (k) of 2 at a confidence interval of 95 % ranged from 7 % to 54 %. The method was applied to the analysis of 44 selected tattoo and PMU ink samples purchased on the Italian market. Phthalate content results were reported taking into account the measurement uncertainty. The uncertainty information is necessary to assess compliance of results with respect to the mass fraction limits established in the upcoming Annex XVII of Regulation (CE) N.1907/2006 (REACH).

DNA/RNA synthesis precursors are especially vulnerable to damage induced by reactive oxygen species occurring following oxidative stress. Guanosine triphosphates are the prevalent oxidized nucleosites, which can be misincorporated during replication, leading to mutations and cell death. Here, we present a novel method based on micro-Raman spectroscopy, combined with ab initio calculations, for the identification, detection, and quantification of low concentrations of oxidized nucleotides. We also show that the Raman signature in the terahertz spectral range (< 100 cm$^{-1}$) contains information on the intermolecular assembly of guanine in tetrads, which allows to further boost the oxidative damage detection limit. Eventually, we provide evidence that similar analyses can be carried out on samples in very small volumes at very low concentrations by exploiting the high sensitivity of Surface Enhanced Raman Scattering combined with properly designed superhydrophobic substrates. These results pave the way for employing such advanced spectroscopic methods for quantitatively sensing the oxidative damage of nucleotides in the cell., 11 pages, 7 figures

Purpose: The mTOR complex C1 (mTORC1) inhibitor everolimus in combination with the aromatase inhibitor exemestane is an effective treatment for patients with hormone receptor—positive (HR+), HER2-negative (HER2−), advanced breast cancer (HR+/HER2− aBC). However, everolimus can cause hyperglycemia and hyperinsulinemia, which could reactivate the PI3K/protein kinase B (AKT)/mTORC1 pathway and induce tumor resistance to everolimus. Experimental Design: We conducted a multicenter, retrospective, Italian study to investigate the impact of baseline and on-treatment (i.e., during first 3 months of therapy) blood glucose levels on progression-free survival (PFS) in patients with HR+/HER2− aBC treated with everolimus-exemestane. Results: We evaluated 809 patients with HR+/HER2− aBC treated with everolimus-exemestane as any line of therapy for advanced disease. When evaluated as dichotomous variables, baseline and on-treatment glycemia were not significantly associated with PFS. However, when blood glucose concentration was evaluated as a continuous variable, a multivariable model accounting for clinically relevant patient- and tumor-related variables revealed that both baseline and on-treatment glycemia are associated with PFS, and this association is largely attributable to their interaction. In particular, patients who are normoglycemic at baseline and experience on-treatment diabetes have lower PFS compared with patients who are already hyperglycemic at baseline and experience diabetes during everolimus-exemestane therapy (median PFS, 6.34 vs. 10.32 months; HR, 1.76; 95% confidence interval, 1.15–2.69; P = 0.008). Conclusions: The impact of on-treatment glycemia on the efficacy of everolimus-exemestane therapy in patients with HR+/HER2− aBC depends on baseline glycemia. This study lays the foundations for investigating novel therapeutic approaches to target the glucose/insulin axis in combination with PI3K/AKT/mTORC1 inhibitors in patients with HR+/HER2− aBC.

The Attirement of the Bride, painted by Max Ernst in 1940, is permanently on display in the Peggy Guggenheim Collection (PGC) in Venice. Within the framework of a larger study of materials and techniques of works by Ernst at the PGC, researchers carried out a series of non-invasive analyses. A combined approach based on Vis-NIR multispectral imaging, Raman Spectroscopy, X-Ray Fluorescence (XRF), and External Reflection Fourier Transform Infrared Spectroscopy (ER-FTIR), was conducted insitu in the museum galleries. In addition, two micro-samples were analysed with SEM-EDS and μ-Raman. This approach produced vital information on the preparatory drawing, the pentimenti, the pigments, the extenders and the alteration products present in the painting. The results showed also the presence of a white preparatory layer, under the pictorial one and above the canvas, composed by lithopone (a mixture of zinc sulphide and barium sulphate), basic lead carbonate and calcium carbonate. The latter compound is probably present also as an extender. The palette included both traditional and new synthetic, commercial pigments, used as primary colours or in mixtures. Moreover, in several areas of the painting, zinc oxalates have been identified as alteration products and more rarely, zinc metal soaps, which could also be present as additives in the paint tube. These analyses revealed the masterful and innovative painting technique of this pioneer of Surrealism.

Breast cancer is the most frequent and lethal tumor in women and finding the best therapeutic strategy for each patient is an important challenge. PARP inhibitors (PARPis) are the first, clinically approved drugs designed to exploit synthetic lethality in tumors harboring BRCA1/2 mutations. Recent evidence indicates that PARPis have the potential to be used both in monotherapy and combination strategies in breast cancer treatment. In this review, we show the mechanism of action of PARPis and discuss the latest clinical applications in different breast cancer treatment settings, including the use as neoadjuvant and adjuvant approaches. Furthermore, as a class, PARPis show many similarities but also certain critical differences which can have essential clinical implications. Finally, we report the current knowledge about the resistance mechanisms to PARPis. A systematic PubMed search, using the entry terms “PARP inhibitors” and “breast cancer”, was performed to identify all published clinical trials (Phase I-II-III) and ongoing trials (ClinicalTrials.gov), that have been reported and discussed in this review.

The airway epithelium contains ionocytes, a rare cell type with high expression of Forkhead Box I1 (FOXI1) transcription factor and Cystic Fibrosis Transmembrane conductance Regulator (CFTR), a chloride channel that is defective in cystic fibrosis (CF). Our aim was to verify if ionocyte development is altered in CF and to investigate the relationship between ionocytes and CFTR-dependent chloride secretion. We collected nasal cells by brushing to determine ionocyte abundance. Nasal and bronchial cells were also expanded in vitro and reprogrammed to differentiated epithelia for morphological and functional studies. We found a relatively high (~3%) ionocyte abundance in ex vivo nasal samples, with no difference between CF and control individuals. In bronchi, ionocytes instead appeared very rarely as previously reported, thus suggesting a possible proximal–distal gradient in human airways. The difference between nasal and bronchial epithelial cells was maintained in culture, which suggests an epigenetic control of ionocyte development. In the differentiation phase of the culture procedure, we used two media that resulted in a different pattern of CFTR expression: confined to ionocytes or more broadly expressed. CFTR function was similar in both conditions, thus indicating that chloride secretion equally occurs irrespective of CFTR expression pattern.

Disposable non-invasive and compatible real time monitoring tool was developed in order to follow the cleaning process of paper artwork directly in situ. This tool was based on a biocompatible cleaning hydrogel coupled with flow electrochemical diagnostic tool, suitable to verify in situ and in a simple way the assessment of degradation of artwork and the efficiency of cleaning process. In this paper, the results obtained by applying this tool on a great format artwork with a lining as support, “Le Nozze di Psiche”, engraved by Diana Scultori, printed in 1613, are reported. This opera was affected by a structural and chromatic deterioration due to a strong oxidative degradation. Such deterioration was probably accelerated by the adhesive (a mixture of starch paste and animal glue) used in a previous lining intervention. In this case, the cleaning agents used are rigid hydrogels of Gellan gum, modified with hydrolytic enzymes. By using the flow sampling system, all materials removed by the gel was carried up to a thin layer cell containing a selective electrochemical biosensors, suitable to monitor both treatments, the cleaning process and the removal of lining. These were monitored, allowing understanding when both processes were completed, thus avoiding lengthy and unnecessary cleaning applications. The effectiveness of cleaning with Gellan gel was assessed quantitatively by using non-invasive optical reflectance spectroscopy by a portable instrumentation, elaborating data with an improved version of the Kubelka-Munk theory in order to recover the absorption coefficient of the cellulose fibers of “Le Nozze di Psiche”. The concentration of oxidized groups acting as chromophores was obtained by comparing the experimental optical absorption spectra to those simulated computationally by using TDDFT-based calculations. By following the cleaning with Gellan gel the results indicate a large decrease of the concentration of degradation product of cellulose. Moreover, chromatographic analysis were carried out in order to evaluate the amount of acid compound, produced during the aging and present on the graphic artwork, using the Gellan gel after cleaning step. The results obtained from the restoration of “Le Nozze di Psiche” have allowed the restorers to evaluate innovative methods for cleaning treatment of paper artworks with a highly specialized scientific-diagnostic approach

The active targeting strategy has emerged as a promising approach to achieve selectivity in nanobiotechnology applications. Peptides are particularly suited as targeting moieties because the multiv...

Campylobacter is an important cause of acute bacterial disease in human worldwide. Within the genus Campylobacter, C. jejuni and C. coli are the two species responsible for over 95% of human infections. The purpose of this work is the development of an Enzyme-Linked-Immuno-Magnetic-Electrochemical (ELIME) assay for a simple and rapid detection of both Campylobacter species. Tosyl-activated magnetic beads (MBs), supporting a sandwich immunological complex, were coupled to a strip of 8-magnetized screen-printed electrodes, as electrochemical transducing element. With the aim to simultaneously detect C. jejuni and C. coli, four anti-campylobacter antibodies were screened in ELISA paying the attention to the selection of both culture medium and cell treatment (whole campylobacter cells inactivated with NaN3 and heat killed cells) for a suitable antigen-antibody interaction. Six combinations of the selected anti-campylobacter antibodies, unconjugated and conjugated with Horseradish Peroxidase (HRP) or Alkaline Phosphatase (AP), were exploited as sensing strategy to set-up the ELIME assay. The antibody pair, consisting of a goat polyclonal antibody anti-campylobacter (coated on the surface of MBs) and the same antibody conjugated with AP (to label the captured cells), was selected given the best response for both Campylobacter species and the lowest detection limit (9 × 103 CFU/mL for C. jejuni and 2 × 104 CFU/mL for C. coli). The selectivity of the ELIME assay was proved by analyzing other C. jejuni strains and non target microorganisms attaining promising results.

The increasing demand by citizens and environmental organization for the protection, preservation, and possible restoration of the marine environment has made seawater protection one of the urgent priorities of the European Union. In this contest, extensive monitoring and surveillance are required to correctly assess the current status of marine environment, paying attention not only to traditional chemical parameters, but also to the so-called “emerging pollutants”. Among these, marine biotoxins represent a serious hazard, because they can accumulate in fish and mussels and enter the food chain. The detection of marine toxins released by algae has mostly been directed on the analysis of fish/shellfish homogenate rather than seawater samples. To fill this gap, in this work, we propose the re-modeling of ELISA kits to set-up non-automated direct competitive ELIMC (Enzyme-Linked Immuno-Magnetic Colorimetric) assays for the detection of Domoic Acid (DA), Okadaic Acid (OA) and Saxitoxin (STX) in seawater. For each toxin, linear working range, sensitivity and detection limit (0.03, 0.05, 0.01 ng/mL for DA, OA and STX, respectively) of the ELIMC assays were calculated. After evaluation of seawater matrix effect and recovery study, ELIMC assays were integrated within a novel automated networked system (ASMAT - Analytical System for Marine Algal Toxins) based on the micro Loop Flow Reactor (μLFR) technology, suitable for on-line monitoring of marine toxins. Once established the best operational conditions, ASMAT was calibrated towards DA, OA and STX verifying its capability to detect sub-ppb levels of the target toxins. Analysis of DA, OA and STX in real samples of marine water, sampled in Alonnisos (Greece) were carried out in laboratory using ELIMC assays, ASMAT and ELISA Kits, for a comparative evaluation. Finally, on-line suitability of ASMAT as an early warning alarm system was demonstrated by a field installation within a floating platform in the port of La Spezia (Italy). © 2018 Elsevier B.V.

This paper reports atomic force microscopy results and molecular dynamics simulations of the striking differences of long-term self-organization structures of negatively charged (AcA4)2KD (double tail) and AcA4D (single tail) peptides, respectively, forming micrometer-long, linearly ordered ribbon-like structures and nanometer-sized, unstructured, round-shaped aggregates. The subsequent formation steps of the long-range nanoribbons, experimentally observed only for the “double tail” (AcA4)2KD peptide, are analyzed in detail, showing that the initial “primary” unstructured round-shaped aggregates progressively evolve into longer nanofilaments and into micrometer-long, network-forming nanoribbon moieties. In particular, the long-range self-organization of the “double tail” peptides appears to be closely related to electrostatically driven diffusional motions of the primary aggregates and nanofilaments. The diffusional freedom degrees are prompted by the formation of a dynamic ternary air/liquid/substrate interface, due to the water evaporation process from the ultrathin films of the peptide solution cast onto a solid mica substrate. Overall, the initial aggregation of unstructured round-shaped moieties, for both the peptides, can be seen as an entropy-driven process, involving the intra- and intermolecular interactions of hydrophobic parts of the peptides, while the further formation of long nanoribbons, only for “double tail” peptides, can be viewed in terms of an enthalpy-driven process, mainly due to the predominant electrostatic interactions between the charged heads of the interacting peptides. The role of the solid–liquid interface, as the locus of the enthalpy-driven linear organization, is also highlighted.

Cell targeting has been considered an important strategy in diagnostic and therapeutic applications. Among different targeting units, peptides have emerged for their ability to bind to many different cellular targets, their scarce immunogenicity and the possibility of introducing multiple copies on nanosystems, providing high avidity for the target. However, their sensitivity to proteases strongly limits their applications in vivo. Here, we show that when presented on the surface of nanostructures, peptide stability to proteolysis is strongly improved without reducing the targeting activity. We prepared plasmonic nanostructures functionalized with a dodecapeptide (GE11) which targets EGFR, a protein overexpressed on different types of tumors. Two types of nanosystems were prepared in which the targeting unit was either directly linked to gold nanoparticles or through a PEG chain, resulting in a different peptide density on the surface of nanostructures. The peptide was rapidly degraded in 20% human serum or in the presence of isolated serine proteases, whereas no significant proteolytic fragments were detected during incubation of the nanosystems and after 24 h digestion, the nanostructures maintained their targeting activity and selectivity on colon cancer cells. Molecular dynamic calculations of the interaction of the nanostructure with chymotrypsin suggest that the formation of the enzyme-peptide complex, the first step in the mechanism of peptide hydrolysis, is highly unlikely because of the constraint imposed by the link of the peptide to the nanoparticle. These results support the utilization of peptides as active targeting units in nanomedicine.

Background: Eribulin shows some activity in controlling brain metastasis in breast cancer. Methods: This observational, multicenter study evaluated brain disease control rates, survival and safety in patients with brain metastatic breast cancer treated with eribulin in clinical practice. Results: A total of 34 patients were enrolled (mean age 49 years, 91% with visceral metastases) and 29 were evaluable for brain disease. Fourteen achieved disease control and showed a longer time without progression: 10 months (95% CI: 2.3–17.7) versus 4 months (95% CI: 3.3–4.7) in the control group (p = 0.029). Patients with clinical benefits at 6 months had longer survival. Leukopenia and neutropenia were the most frequent grade 3–4 toxicities. Conclusion: Eribulin confirms its effectiveness in patients with brain metastatic breast cancer. Further studies on larger cohorts are needed to confirm the results.

In this work, we shed new light on ultrasound contrast agents applied to the field of cultural heritage as an invaluable fine-tune cleaning tool for paper artworks. In this context, one of the primary and challenging issues is the removal of modern adhesives from paper artifacts. Modern adhesives are synthetic polymers whose presence enhances paper degradation and worsens its optical features. A thorough analytical and high-spatial-resolution combined study was successfully performed to test the capability of poly(vinyl alcohol)-based microbubbles stimulated by a proper noninvasive 1 MHz ultrasound field exposure in removing these adhesives from paper surfaces, in the absence of volatile invasive and toxic chemicals and without damaging paper and/or leaving residues. We demonstrate that poly(vinyl alcohol)-shelled microbubbles are suitable for interacting with paper surfaces, targeting and boosting in a few minutes the nondamaging removal of adhesive particles from paper samples thanks to their peculiar shell composition together with their ultrasound dynamics.

An analytical methodology based in the combination of Thin Film Microextraction with Laser-induced Breakdown Spectroscopy (TFME-LIBS) was investigated, for the first time, for detection of Cu, Cr, Ni and Pb in aqueous solutions. In this methodology, the analytes were extracted in a thin film of adsorbent material deposited on a solid support, which was introduced in the sample to analyse. After extraction, the analytes retained in the adsorbent were analysed by LIBS. In order to obtain adsorbent films useful for the microextraction step, two different experimental procedures for film generation, denoted as Drop Casting Deposition and Mould Deposition, were evaluated. In both cases, graphene oxide was used as adsorbent material. The mould deposition procedure was found to produce more homogeneous graphene oxide layers, leading to more uniform distribution of the adsorbed analytes on the graphene oxide surface. Experimental parameters affecting the TFME procedure, such as the adsorbent amount and extraction time, were studied. Under optimum microextraction conditions, the analytical figures of merit of the proposed TFME-LIBS method were evaluated, leading to limits of detection ranging from 41 μg kg−1 and 52 μg kg−1. Method trueness, evaluated from the analysis of a real sample of bottle water, led to recovery values about 70%, indicating the existence of strong matrix effects probably due to the presence of major cations in the bottle water. After 50% dilution of the sample with deionized water, recoveries values improved to 100%–108%. This work was supported by the Spanish Ministry of Economy and Competitiveness [CTQ2016-79991-R], the Regional Government of Valencia (Spain) [PROMETEO/2013/038 and PROMETEO/2018/087]; and the University of Alicante [UAUSTI17-04 and UAUSTI18-04]. L. Ripoll is grateful to the Ministry of Economy and Competitiveness for her PhD fellowship (FPI-MICINN (BES-2012-058759)).

Doping profiling methods are required to provide doping monitoring of heavy doped Silicon wafers, widely used in electronic devices, with high concentration sensitivity and spatial resolution. Herein, we demonstrate that ultraviolet (UV) micro-Raman spectroscopy implemented on small-angle beveled surfaces is able to produce a Raman-based doping profile without any further measurements. The reliability of the approach is verified on ion-implanted p-type B-doped single-crystalline Si (100) wafers with shallow (100 nm) doping profile, directly comparing the Raman-based doping profiling results with independent secondary ion mass spectrometry (SIMS) and spreading resistance profiling (SRP) measurements. The Raman-based technique is capable to fully reproduce the doping profile down to 100 nm with excellent doping sensitivity (10 ppm) in the 1018–1020 cm−3 doping concentration range providing high lateral (vertical) resolution of 1 μ m (10 nm). The technique is comparable to SIMS in terms of resolution and sensitivity down to 1018 cm−3 concentration but requires the beveling process. Instead, it is a better alternative to SRP on bevel due to its higher spatial resolution and avoiding probe contact. The Raman-based doping profiling technique is, therefore, suitable for heavy doped Si with typical shallow doping profiles of state-of-the-art Si-based nanoelectronics.

Herein, we propose the first three-dimensional origami paper-based device for the detection of several classes of pesticides by combining different enzyme-inhibition biosensors. This device was developed by integrating two different office paper-based screen-printed electrodes and multiple filter paper-based pads to load enzymes and enzymatic substrates. The versatile analysis of different pesticides was carried by folding and unfolding the filter paper-based structure, without any addition of reagents and any sample treatment (i.e. dilution, filtration, pH adjustment). The paper-based platform was employed to detect paraoxon, 2,4-dichlorophenoxyacetic acid, and atrazine by exploiting the capability of these different types of pesticides (i.e. organophosphorus insecticides, phenoxy-acid herbicides, and triazine herbicide) to inhibit butyrylcholinesterase, alkaline phosphatase, and tyrosinase, respectively. The degree of inhibition correlating to the quantity of pesticides was evaluated by chronoamperometrically monitoring the enzymatic activity in the absence and in the presence of pesticides by using a portable potentiostat. To improve the sensitivity, the paper-based electrodes were modified with carbon black alone in the case of platforms for 2,4-dichlorophenoxyacetic acid and atrazine detection, or decorated with Prussian blue nanoparticles for the detection of paraoxon. The paper-based device was applied for the detection of paraoxon, 2,4-dichlorophenoxyacetic acid, and atrazine at ppb level in both standard solutions and river water sample. The accuracy of this origami multiple paper-based electrochemical biosensor was evaluated in river water by recovery studies, obtaining satisfactory values (e.g. for paraoxon 90 ± 1% and 88 ± 2%, for 10 and 20 ppb, respectively). The proposed three-dimensional origami paper device allows for rapid, cost-effective and accurate pesticide detection in surface water as a result of combining filter and office papers, screen-printing, wax-printing and nanomaterial technology.

CREKA (Cys–Arg–Glu–Lys–Ala) and its engineered analogue CRMeEKA, in which Glu has been replaced by N-methyl-Glu to provide resistance against proteolysis, are emerging pentapeptides that were specifically designed to bind fibrin–fibronectin complexes accumulated in the walls of tumour vessels. However, many of the intrinsic properties of CREKA and CRMeEKA, which are probably responsible for their different behaviour when combined with other materials (such as polymers) for diagnosis and therapeutics, remain unknown yet. The intrinsic tendency of these pentapeptides to form aggregates has been analysed by combining experimental techniques and atomistic Molecular Dynamics (MD) simulations. Dynamic light scattering assays show the formation of nanoaggregates that increase in size with the peptide concentration, even though aggregation occurs sooner for CRMeEKA, independently of the peptide concentration. FTIR and circular dichroism spectroscopy studies suggest that aggregated pentapeptides do not adopt any secondary structure. Atomistic MD trajectories show that CREKA aggregates faster and forms bigger molecular clusters than CRMeEKA. This behaviour has been explained by stability of the conformations adopted by un-associated peptide strands. While CREKA molecules organize by forming intramolecular backbone – side chain hydrogen bonds, CRMeEKA peptides display main chain – main chain hydrogen bonds closing very stable ¿- or ß-turns. Besides, energetic analyses reveal that CRMeEKA strands are better solvated in water than CREKA ones, independent of whether they are assembled or un-associated.

Wet cleaning of paper artworks by using hydrogels is a recently proposed process in the cultural heritage field. In this contest, on one side, research has focused on more effective and tuned cleaning hydrogels; on the other, the cleaning performances of the proposed hydrogels have been studied on ancient paper and very little on modern paper. This lack of information is not negligible, since the two kinds of paper differ strongly in composition. The difference, in turn could affect the performance of cleaning gels. In this article, new chemical hydrogels were synthetized using an oxidized polyvinyl alcohol (PVA) bearing an aldehyde group at each chain end, as cross-linking agent, i.e. telechelic PVA. PVA hydrogels have been characterized as candidates for paper cleaning applications. The possibility of tuning the mechanical and retentive properties, simply by changing the ratio of two polymer concentrations, their stability and transparency, indeed, render them, potentially suitable in cultural heritage field. The cleaning capability of two hydrogels, with different PVA/telechelic PVA ratios, has been assessed both on ancient paper, than a modern one and the results have been compared. The efficacy of these hydrogels has been demonstrated characterizing the samples before and after the cleaning process by means of a multidisciplinary approach involving spectroscopic and chromatographic tests.

For reproducing the behavior of water molecules adsorbed on gold surfaces in terms of density of both bulk and interfacial water and in terms of structuring of water on top of gold atoms, the implementation of a multibody potential is necessary, thus the Stillinger–Weber potential was tested. The goal is using a single nonbonded potential for coarse-grained models, without the usage of explicit charges. In order to modify the angular part of the Stillinger–Weber potential from a single cosine to a piecewise function accounting for multiple equilibrium angles, employed for Au–Au–Au and Au–Au–water triplets, it is necessary to create a version of the simulation package LAMMPS that supports the assignment of multiple favored angles. This novel approach is able to reproduce the data obtained using quantum mechanical calculations and density profiles of both bulk and adsorbed water molecules obtained using classical polarizable force fields. published

Giulio Turcato (Mantova 1912–Rome 1995) was an Italian artist, belonging to the figurative and abstract expressionist currents. At the end of the 1960s, fascinated by the US space program, Turcato produced a series of artworks, differing in colour and shape, titled Superficie Lunare (Moon Surface). To reproduce a realistic moon surface, Turcato used polyurethane foam as support for the painting. The combination of this unconventional material and the pictorial ones makes Superficie Lunare an interesting case of study to understand the alteration phenomena in contemporary art. In this study, a blue painted Superficie Lunare (1969) was analysed by means of non-invasive (ER-FTIR, Raman and X-ray spectroscopy) and micro-invasive techniques (µ-ATR, μ-FTIR, μ-Raman, Py-GC/MS, EGA-MS and SEM-EDS). The results point out the use of a polyurethane foam as support and of a poly(vinyl acetate) based Prussian blue paint tube as main constituent of the paint layer. The analyses highlight also the complexity of the paint layer, showing the presence of lampblack and Al foil mixed to the blue paint, lithopone inside the white crater and possibly ferric oxide in the red one. The results confirm the use of a polyurethane foam for the support. The painted layers present a binder composed of poly(vinylacetate), with polystyrene, possibly present as additive. Concerning the nature of the blue pigment, Prussian blue was identified on the whole surface. In some points, also lampblack was detected. Furthermore, small foils of Al were found inside the paint layer, probably to give sparkling effect. Inside the moon craters, lithopone was identified for the white pigment and possibly iron oxide for the red one. The multi-technique approach allowed also to obtain information about the manufacturing of the used Prussian blue and the occurrence of alteration process for the polyurethane foam.

In this work, we report the first application of the double-pulse Calibration-Free Laser-Induced Breakdown Spectroscopy (DP-CF-LIBS) to the analysis of Ge-Cu/Si and Ge-Ba/Si alloys. The plasma was produced using two collinear Nd:YAG lasers, operating at 1064 nm. The experimental parameters such as inter-pulse delay, gate delay and energy ratio between the two laser pulses were optimized to maximize the signal to background and signal to noise ratio in the resulting LIBS spectra. The fulfillment of the Local Thermodynamic Equilibrium (LTE) approximation in the time window of the measurement was verified using the McWhirter criterion and considering the relaxation time and diffusion length of the atoms in the plasma. The effect of self-absorption on the lines used in the CF-LIBS analysis was considered and found negligible. The DP-CF-LIBS analysis allowed the determination of the stoichiometric ratio of the elements in the two alloys, an important parameter for industrial quality control and fundamental studies on germanium/silicon alloys.

Fine airborne particulate matter (PM2.5) has been repeatedly associated with adverse health effects in humans. The PM2.5 soluble fraction, and soluble metals in particular, are thought to cause lung damage. Literature data, however, are not consistent and the role of leachable metals is still under debate. In this study, Winter and Summer urban PM2.5 aqueous extracts, obtained by using a bio-compatible solution and different contact times at 37 °C, were used to investigate cytotoxic effects of PM2.5 in cultured lung epithelial cells (A549) and the role played by the leachable metals Cu, Fe, Zn, Ni, Pb and Cd. Cell viability and migration, as well as intracellular glutathione, extracellular cysteine, cysteinylglycine and homocysteine concentrations, were evaluated in cells challenged with both PM2.5 extracts before and after ultrafiltration and artificial metal ion solutions mimicking the metal composition of the genuine extracts. The thiol oxidative potential was also evaluated by an abiotic test. Results demonstrate that PM2.5 bioactive components were released within minutes of PM2.5 interaction with the leaching solution. Among these are i) low MW (

Chemical warfare agents (CWAs) have become a crucial concern of the 20th century in the safety vision of the society and the environment, triggering a growing awareness for early alarm in case of terroristic attacks as well as preventing contamination. Herein, we present the development of a whole-cell optical bioassay for the detection of CWA simulants. In particular, the effects of the mustard agent simulants bis-2-chloroethyl amine and 2-chloroethyl ethyl sulphide on the unicellular green photosynthetic algae Chlamydomonas reinhardtii was studied, with the aim to unravel the response of the microorganism to the presence of these simulants and optimise the analytical conditions of the bioassay. Important variations in the growth, photosynthetic activity, and content of photosynthetic pigments were observed in the presence of the selected simulants. The algal response towards bis-2-chloroethyl amine and 2-chloroethyl ethyl sulphide in a concentration range between 0.2 and 2.5 mM was analysed, indicating a linear relationship in the measured dose-response curves and detection limits of 50 and 200 μM, respectively. Interference studies demonstrated the suitability of the proposed optical bioassay to detect mustard agent simulants also in drinking water, a defenceless matrix in case of terroristic attack, where atrazine, copper, and arsenic could be present at safety limits. Very slight matrix effect was evidenced, with ratios of 1.04 and 0.86 for calibration curves of bis-2-chloroethyl amine and 2-chloroethyl ethyl sulfide in tap water samples with respect to curves of standard solutions. Recovery values of 104 ± 15% and 97.5 ± 6.5% for 1 mM and 2 mM of bis-2-chloroethyl amine and 93 ± 16% and 105 ± 5% for 1 mM and 2 mM of 2-chloroethyl ethyl sulfide were achieved.

When compared to other analytical techniques, LIBS shows relatively low precision and, generally, high Limits of Detection (LODs). Until recently, the attempts in improving the LIBS performances have been based on the use of more stable/powerful lasers, high sensitivity detectors or controlled environmental parameters. This can hinder the competitiveness of LIBS by increasing the instrumental setup cost and the difficulty of operation. Sample treatment has proved to be a viable and simple way to increase the LIBS signal; in particular, the Nanoparticle-Enhanced Laser Induced Breakdown Spectroscopy (NELIBS) methodology uses a deposition of metal nanoparticles on the sample to greatly increase the emission of the LIBS plasma. In this work, we used a simple, fast, “green” and low-cost method to synthetize silver nanoparticles by using coffee extract as reducing agents for a silver nitrate solution. This allowed us to obtain nanoparticles of about 25 nm in diameter. We then explored the application of such nanoparticles to the NELIBS analysis of metallic samples with a mobile LIBS instrument. By adjusting the laser parameters and optimizing the sample preparation procedure, we obtained a NELIBS signal that is 4 times the LIBS one. This showed the potential of green-synthetized nanoparticle for NELIBS applications and suggests the possibility of an in-situ application of the technique.

Experimental and theoretical reports have shown that nanostructured surfaces have a dramatic effect on the amount of protein adsorbed and the conformational state and, in turn, on the performances of the related devices in tissue engineering strategies. Here we report an innovative method to prepare silica-based nanostructured surfaces with a reproducible, well-defined local curvature, consisting of ordered hexagonally packed arrays of curved hemispheres, from nanoparticles of different diameters (respectively 147 nm, 235 nm and 403 nm). The nanostructured surfaces have been made chemically homogeneous by partially embedding silica nanoparticles in poly(hydroxymethylsiloxane) films, further modified by means of UV-O3 treatments. This paper has been focused on the experimental and theoretical study of laminin, taken as a model protein, to study the nanocurvature effects on the protein configuration at nanostructured surfaces. A simple model, based on the interplay of electrostatic interactions between the charged terminal domains of laminin and the nanocurved charged surfaces, closely reproduces the experimental findings. In particular, the model suggests that nanocurvature drives the orientation of rigid proteins by means of a "geometrical resonance" effect, involving the matching of dimensions, charge distribution and spatial arrangement of both adsorbed molecules and adsorbent nanostructures. Overall, the results pave the way to unravel the nanostructured surface effects on the intra- and inter-molecular organization processes of proteins.

In this study, We demonstrate mono and few layers MoS2 samples on the SiO2(270nm)/Si substrate from bulk MoS2 crystal by micromechanical exfoliation technique. We have systematically studied Atomic Force Microscopy, Raman and PL properties of mono and few layer MoS2 on the SiO2(270nm)/Si substrate. First, we find that the number of layer values dependent the Raman and PL emission. First, Raman intensity area ratio of the MoS2E12g, A1g and 2LA modes to that area of the Si substrate increased linear with increasing number of layers MoS2. Second, Normalized PL intensity area of the (A) peak decreased linear with increasing number of layers MoS2. The value of those graphs is a method to understand the number of layers the exfoliated MoS2.

A detailed characterization of the behavior of the amphiphilic antimicrobial peptide Trichogin GA IV (TRIC) at the air/water interface during a Langmuir–Blodgett (LB) isotherm is reported. By means of molecular dynamics simulations, experimental data are explained in terms of the conformational changes and aggregate features adopted by TRIC. We show that, due to compression, different structural changes occur: initially formed drop-like aggregates coalesce, forming nanofibers; on increasing the surface tension further, these nanofibers constitute a web-like structure in which meshes are filled by water pools. During these transitions, the peptide chains lie almost parallel to the surface mostly adopting a helical conformation. At high peptide concentration, reaching the maximum of the allowed surface pressure, a monolayer of TRICs in nonhelical conformation and vertically aligned with respect to the air/water interface is formed.

The rapidly expanding scenario of treatment options for patients affected by prostate cancer (PC) is leading to improved outcomes; however, PC still represents one of the most frequent causes of male mortality. Thus, while translational research is trying to unravel the molecular landscape underlying carcinogenesis, disease progression and treatment resistance, several clinical trials are evaluating novel options to further expand therapeutic options. The cyclin-dependent kinases (CDK)-pathway represents a promising therapeutic target for different cancer types; due to the pivotal role of this pathway in the regulation of PC cell cycle, three CDK4/6-inhibitors (abemaciclib, palbociclib and ribociclib) are currently being investigated in several clinical trials. In this paper, we review the current knowledge on CDK-pathway and the mechanism of action of CDK-inhibitors; we discuss the biological rationale for their use in PC and the state of the art of clinical trials focused on the demonstration of their potential role in early or advanced stage, in hormone-sensitive and castration-resistant state. Finally, the potential application of precision oncology for treatment selection in PC is discussed.

An Enzyme Linked ImmunoMagnetic Electrochemical assay (ELIME) was developed for the detection of the hepatitis A virus (HAV). This system is based on the use of new polydopamine-modified magnetic nanobeads as solid support for the immunochemical chain, and an array of 8 screen-printed electrodes as a sensing platform. Enzymatic-by-product is quickly measured by differential pulse voltammetry. For this purpose, all analytical parameters were optimized; in particular, different blocking reagents were evaluated in order to minimize the nonspecific interaction of bioreagents. Using the ELIME assays, a quantitative determination of HAV can be achieved with a detection limit of 1·10−11 IU mL−1 and a working range between 10−10 – 5 × 10−7 IU mL−1. The cross-reactivity of the commercial monoclonal antibodies against HAV used in ELIME assays was tested for Coxsackie B4, resulting very low. The sensitivity was also investigated and compared with spectrophotometric sandwich ELISA. The average relative standard deviation (RSD) of the ELIME method was less than 5% for the assays performed on the same day, and 7% for the measurements made on different days. The proposed system was applied to the cell culture of HAV, which title was quantified by Real-Time Quantitative Reverse Transcription PCR (RT¬qPCR). To compare the results, a correlation between the units used in ELIME (IU mL−1) and those used in RT¬qPCR (genome mL−1) was established using a HAV-positive sample, resulting in 1 IU mL−1–10−4 gen mL−1 (R2 = 0.978). The ELIME tool exhibits good stability and high biological selectivity for HAV antigen detection and was successfully applied for the determination of HAV in tap water.

The calcium‑phosphorus ratio (Ca/P) has an important role in calcium phosphate applications, as a slight change in this ratio results in substantial modifications of their characteristics. Hydroxyapatite is a bioactive ceramic of the calcium phosphate family, and its composition and crystalline structure are similar to the human bone. For this reason, it is widely used to promote bone integration in prosthetics and scaffolds. Prosthetics are usually made of metal and coated with bioceramics and the some techniques used to make these coatings depositions conserve the stoichiometry from the target during deposition. Therefore, studying the target Ca/P is a way to guaranteeing the final deposition stoichiometry. In this study, we investigated the One Point Calibration – Laser Induced Breakdown Spectroscopy (OPC-LIBS) method for the measurement of the Ca/P of hydroxyapatite targets. This method is quantitative, based on the determination of correction parameters via analysis of a standard sample with known stoichiometry. Results were compared to well-established techniques of X-ray Fluorescence (XRF) and Atomic Absorption Spectroscopy (AAS) and showed that the deviation in the difference is less than 5%, proposing a substitute for elemental analysis. Moreover, this method has technical advantages such as rapid measurements, no need for sample preparation or destruction and easy automation.

The doping concentration of B doped single-crystal Czochralski Si(1 0 0) wafers (6 × 1014-5 × 1019 cm−3) has been monitored via micro-Raman spectroscopy using visible (633 and 532 nm) and near-UV (355 nm) laser excitations at low power (5 mW). Data have been analysed with unprecedented accuracy via a convoluted Fano-Gaussian model of the first-order Raman Stokes mode of Silicon. This allowed the determination of the fitting spectral parameters (peak position and width) with an accuracy of 0.01 cm−1, which enables a reliable probing of the concentration. We observed, independently on the excitation wavelength used, a widening (up to 6.5 cm−1), a frequency-softening (up to 1.5 cm−1) and an intensity reduction (down to 90 % ) of the Si peak with the doping concentration. The widening and frequency-softening follow a strictly linear dependence with doping concentration, allowing a calibration. A linear dependence of the reciprocal Fano asymmetry parameter ( q - 1 ) with excitation energy is verified, with the slope showing a linear behavior with the doping concentration and providing a direct estimate on the hole-phonon interaction strength. Results are reproduced with surface-sensitive near-UV Raman spectroscopy on BF 2 + ion implanted and laser thermal annealed (LTA) Si, demonstrating the full portability of the Raman technique to state-of-the-art nanoelectronics.

Herein we describe a novel reagentless paper-based electrochemical phosphate sensor, manufactured with a simple and inexpensive approach. By following three easy steps, consisting of wax patterning, paper chemical modification, and electrode screen-printing, the filter paper provides an effective electroanalytical platform to sense phosphate ions in standard solutions and real samples (river water). The electrochemical properties of the paper-based platform were evaluated, firstly, by using ferricyanide as a redox mediator, proving no analyte-entrapment due to the cellulose lattice. Then, the reference colorimetric method for phosphate ions, which is based on the formation of phosphomolybdic complex, was successfully adapted to a reagentless electrochemically paper-based platform. This novel and highly sustainable configuration readily allows for the determination of phosphate ions with high reproducibility and long storage stability, achieving a detection limit of 4 μM over a wide linear range up to 300 μM. This in-house approach would be able to generically develop an affordable in situ and user-friendly sensing device without the addition of any reagent, to be applied for a broad range of analytes.

International audience; Proton secretion mediated by ATP12A protein on the surface of the airway epithelium may contribute to cystic fibrosis (CF) lung disease by favoring bacterial infection and airway obstruction. We studied ATP12A in fresh bronchial samples and in cultured epithelial cells. In vivo, ATP12A expression was found almost exclusively at the apical side of nonciliated cells of airway epithelium and in submucosal glands, with much higher expression in CF samples. This could be due to bacterial infection and inflammation, since treating cultured cells with bacterial supernatants or with IL-4 (a cytokine that induces goblet cell hyperplasia) increased the expression of ATP12A in nonciliated cells. This observation was associated with upregulation and translocation of ATP1B1 protein from the basal to apical epithelial side, where it colocalizes with ATP12A. ATP12A function was evaluated by measuring the pH of the apical fluid in cultured epithelia. Under resting conditions, CF epithelia showed more acidic values. This abnormality was minimized by inhibiting ATP12A with ouabain. Following treatment with IL-4, ATP12A function was markedly increased, as indicated by strong acidification occurring under bicarbonate-free conditions. Our study reveals potentially novel aspects of ATP12A and remarks its importance as a possible therapeutic target in CF and other respiratory diseases.

Bioassays based on enzyme inhibition are analytical tools widely employed for inhibitor analysis. Beside the conventional analytical techniques such as chromatography and mass spectrometry, these bioassays are cost-effective, easy to use, and suitable for in situ measurement but they are often characterised by a quite narrow linear range. Herein, we report a novel graphical method based on integrated Michaelis-Menten equation, valid for all types of reversible inhibition, which provides an extended linear range. The suitability of this innovative approach was demonstrated in the case of fluoride quantification using a colorimetric bioassay based on acetylcholinesterase inhibition. The “half time reaction”, estimated by the progress curve of cholinesterase inhibition, was plotted versus the fluoride inhibitor concentration, observing an extended linear range up to 5 mM, instead of 0.6 mM using initial rate measurements. The applicability of this new concept was further demonstrated in the case of catalase enzyme inhibited by cyanide. Furthermore, it was demonstrated that fixed substrate conversion at level of 10–50% allows determination of inhibitor concentration in a wide linear range with high precision and in short time of analysis. This novel theoretical and practical approach allows for the extension of the linear range without any further experiments, with several advantages including low reagent consumption, reduced waste generation and time of measurement.

A dodecapeptide (AMRKLPDAPGMH) functionalized with a tetramethylrhodamine (TAMRA) chromophore at the N-terminus was immobilized on nanocrystalline TiO2. The optical and binding properties of the peptide layer immobilized on the titania surface were characterized by UV–vis absorption, steady-state fluorescence and X-ray photoelectron spectroscopies. Circular Dichroism experiments and Molecular Mechanics calculations showed that the predominant conformation populated by the peptide scaffold brings Arg3, Lys4 and Asp7 in the correct position for linking the TiO2 surface. Photocurrent generation experiments were therefore carried out to determine the photon-to-current conversion efficiency (IPCE) of a Grätzel-like Dye Sensitized Solar Cell (DSSC), the photoactive unit of which is formed by TAMRA-AMRKLPDAPGMH/TiO2. The measured IPCE amounted to 0.65%, a value that is definitely low, but superior to those previously reported for similar bioinspired DSSCs. This result can be ascribed to the light-harvesting properties of the TAMRA chromophore and to the unique structural properties of the peptide spacer. © 2017 Elsevier B.V.